Assessing mitochondrial morphology and dynamics using fluorescence wide-field microscopy and 3D image processing.
Mitochondrial morphology and length change during fission and fusion and mitochondrial movement varies dependent upon the cell type and the physiological conditions. Here, we describe fundamental wide-field fluorescence microscopy and 3D imaging techniques to assess mitochondrial shape, number and length in various cell types including cancer cell lines, motor neurons and astrocytes. Furthermore, we illustrate how to assess mitochondrial fission and fusion events by 3D time-lapse imaging and to calculate mitochondrial length and numbers as a function of time. These imaging methods provide useful tools to investigate mitochondrial dynamics in health, aging and disease.
Song
Wenjun
W
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.
Bossy
Blaise
B
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.|The Burnham Institute for Medical Research, 10901 N. Torrey Pines Road, La Jolla, CA 92037.
Martin
Ola J
OJ
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.
Hicks
Andrew
A
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.|University of Pennsylvania, 3451 Walnut Street, Philadelphia, PA.
Lubitz
Sarah
S
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.
Knott
Andrew B
AB
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.
Bossy-Wetzel
Ella
E
Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, 4000 Central Florida Blvd., Orlando, FL 32816.|The Burnham Institute for Medical Research, 10901 N. Torrey Pines Road, La Jolla, CA 92037.
D000818:Animals; D001253:Astrocytes / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008928:Mitochondria / Q000502:physiology / Q000648:ultrastructure*; D053142:Mitochondrial Size; D009046:Motor Neurons / Q000648:ultrastructure; D051381:Rats